There is a need for new technologies to enable the analysis of the protein coding content of a genome. We propose to develop a novel technology for generation and screening of protease substrates. The technology is designed to enable large scale protease activity profiling studies. PUBLIC HEALTH RELEVANCE: The protease activity profiling technology we are developing will help to enable large-scale studies of proteases. This is useful for identifying new drug targets and for drug development. It would help to guide experiments to elucidate the functional roles of specific proteases and may also prove to be a sensitive diagnostic tool, and therefore may result in medical advances in the diagnosis and treatment of disease.